Ultrasound-mediated sonoporation is a promising technique that temporarily permeabilizes cell membranes to enhance delivery of therapeutic agents directly to tumor sites while minimizing systemic side effects. Calcium, a critical regulator of cell death and proliferation, can be introduced into cells by ultrasound, offering a novel therapeutic approach. This study investigates calcium sonoporation (CaSP), which combines ultrasound with calcium ions and microbubbles, to target pediatric rhabdomyosarcoma A204 and RD cells. Our findings showed that CaSP disrupted cellular homeostasis by facilitating the controlled influx of calcium, leading to oxidative stress, mitochondrial dysfunction, cell cycle arrest and activation of apoptotic pathways. The study revealed that the TP53 mutational status significantly influences the cellular response to CaSP. TP53-wild-type A204 cells were particularly susceptible to CaSP, exhibiting marked increases in apoptosis and oxidative damage. In contrast, TP53-mutated RD cells exhibited a reduced oxidative stress and apoptotic response, highlighting the critical role of TP53 in mediating the effects of CaSP. This differential response underscored the potential of TP53 gene as a biomarker for predicting the efficacy of CaSP, offering a pathway toward more personalized cancer therapies. Furthermore, the study demonstrated that CaSP can selectively target cancer cells while sparing healthy tissue. The research laid the groundwork for future studies to optimise sonoporation parameters and explore its integration with existing cancer treatments. The insights gained from this study pave the way for developing more personalized cancer treatment strategies, particularly for tumors influenced by specific genetic contexts, such as TP53 mutations.